Development of a qPCR for the detection of infectious laryngotracheitis virus (ILTV) based on the gE gene

Autor(es)

Parra, Silvana Santander ^[1] ; Nunez, Luis ^[1] ; Buim, Marcos R. ^[2] ; Astolfi-Ferreira, Claudete S. ^[1] ; Piantino Ferreira, Antonio J. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Sch Vet Med, Dept Pathol, Ave Prof Dr Orlando M Paiva 87, BR-05508270 Sao Paulo, SP - Brazil

^[2] Inst Biol, Lab Avian Dis, Bastos, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: BRITISH POULTRY SCIENCE; v. 59, n. 4, p. 402-407, 2018.
• Citações Web of Science: 0

Resumo

1. Infectious laryngotracheitis is a respiratory disease that affects the poultry industry worldwide. It is common in flocks with high-bird density, causing major economic losses.2. In this study, a SYBR (R) FAST polymerase chain reaction (PCR) double-strand DNA intercalating agent assay was performed for the detection of infectious laryngotracheitis virus (ILTV) in clinical samples in comparison with a conventional nested-PCR, both based on the glycoprotein E encoding gene. This assay amplified 56bp and was capable of detecting 1(9) to 1 copies of virus.3. In total, 164 clinical samples were obtained from birds with respiratory problems from the period of 2009-2016. In the nested-PCR, there were 45.12% positive samples and 54.88% negative samples, while in the real-time PCR (qPCR), there were 81.1% positive samples and 18.9% negative samples.4. In conclusion, qPCR from the DNA double-strand intercalating agent SYBR (R) GREEN FAST was useful for the diagnosis of ILTV because it detected samples that were negative in nested-PCR. This assay has advantages, such as a shortened processing-time, and no need for post-amplification processing (electrophoresis) with additional reagents, such as MgCl2 and agarose. Hence, qPCR proved to be useful, rapid and low cost for use with clinical samples. (AU)

• Processo FAPESP: 12/20912-1 - Estudo da prevalência, caracterização molecular e das interações entre as micoplasmoses aviárias e os vírus de laringotraqueíte infecciosa das aves, em poedeiras da região de Bastos
• Beneficiário: Marcos Roberto Buim
• Modalidade de apoio: Auxílio à Pesquisa - Regular